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Chelex 100 as a Medium for Simple Extraction of DNA for PCR-Based Typing from Forensic Material

机译:Chelex 100作为简单提取DNA的介质,用于从法医材料中进行基于PCR的打字

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摘要

Procedures utilizing Chelex100 chelating resin have been developed for extracting DNA from forensic-type samples for use with the PCR. The procedures are simple, rapid, involve no organic solvents and do not require multiple tube transfers for most types of samples. The extracion of DNA from semen and very small blood stains using Chelex 100 is as efficient or more efficient than using proteinase K and phenol-chloroform extraction. DNA extracted from bloodstains seems less prone to contain PCR inhibitors when prepared by this method. The Chelex method has been used with amplification and typing at the HLA DQ alpha locus to obtain the DQ alpha genotypes of many different types of samples, including whole blood, bloodstains, seminal stains, buccal swabs, hair and post-coital samples. The results of a concordance study are presented in which the DQ genotypes of 84 samples prepared using Chelex or using conventional phenol-chloroform extraction are compared. The genotypes obtained using the two different extraction methods were identical for all samples tested.
机译:已经开发了利用Chelex100螯合树脂的方法,可从法医型样品中提取DNA并用于PCR。该过程简单,快速,不涉及有机溶剂,并且对于大多数类型的样品都不需要多次转移试管。与使用蛋白酶K和苯酚-氯仿提取相比,使用Chelex 100从精液和非常小的血迹中提取DNA效率更高或更有效。用这种方法制备时,从血迹中提取的DNA似乎不太可能含有PCR抑制剂。 Chelex方法已用于HLA DQα基因座的扩增和分型,以获得许多不同类型样品的DQα基因型,包括全血,血迹,精液染色,口腔拭子,头发和性交后样品。给出了一致性研究的结果,其中比较了使用Chelex或使用常规酚-氯仿萃取制备的84个样品的DQ基因型。使用两种不同提取方法获得的基因型对于所有测试样品均相同。

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